XENOBIOTIC METABOLISM IN CRYOPRESERVED LIVER SLICE

冷冻保存的肝切片中的异生物代谢

• 批准号: 2154866
• 负责人: KLAUS BRENDEL
• 金额: $ 18.68万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2154866
• 关键词: DNA; adenosine triphosphate; alternatives to animals in research; cryopreservation; cytochrome P450; drug metabolism; enzyme activity; glucuronates; glucuronosyltransferase; growth media; hepatotoxin; human tissue; isozymes; liver metabolism; organ culture; oxygen tension; potassium; protein biosynthesis; secretion; sulfates; temperature; toxin metabolism

The goal of this proposal is to establish the feasibility of human liver slice cultures for the study of xenobiotic metabolism. This new in vitro system has been extensively tested in our laboratory and others with tissue slices from other species and using other end points then xenobiotic metabolism. The novel aspect of this proposal is the application of this prior experience to human tissue and the thorough optimization of all connected parameters such that a powerful new tool, which will find its place in mechanistic human hepatotoxicity studies is made available to the scientific community. Since human tissues are not conveniently available at any time it is important to investigate storage modalities such as banking of cryopreserved slices under liquid nitrogen and cold preservation in special tissue preservation solutions at just above 0 degrees C. Our limited amount of preliminary information seems to indicate that cryopreservation and cold preservation of human tissue slices can be optimized such that metabolic activities of fresh tissue are retained. Through systematic evaluation of the relevant parameters in cryo and cold preservation such as optimal composition of solutions, freezing rate, cryoprotectants vitrification conditions, rewarming rates and their effect on the maintenance of xenobiotic metabolism by the human tissue much needed general knowledge on this new in vitro tool in biochemical pharmacology will emerge. The stability of xenobiotic metabolic pathways during culture of human liver and kidney slices at quasi physiological conditions will teach what can maximally be expected from this in vitro system. Since the focus of this proposal is integrated xenobiotic metabolism i.e. the tight coupling of phase I and II under the conditions of intermediary metabolic control of an intact cellular system we will not attempt at this time to determine amounts, maximal activities, rates of synthesis and degradation of individual enzymes, but will focus on complex metabolite patterns of a few standard xenobiotics and their shifting appearance as individual enzymes drop out or emerge during culture.

该提案的目标是建立人类肝脏的可行性 用于研究外源代谢的切片培养物。 这种新的体外 系统已在我们的实验室和其他实验室进行了广泛的测试 来自其他物种的组织切片并使用其他终点 外源代谢。 该提案的新颖之处在于 将先前的经验应用于人体组织并彻底 优化所有连接参数，从而形成强大的新工具， 它将在人类肝毒性机制研究中占有一席之地 提供给科学界。 由于人体组织不 随时可用，调查存储很重要 液氮下冷冻保存切片等方式 并在特殊的组织保存溶液中冷藏 高于 0 摄氏度。我们有限的初步信息似乎 表明人体组织的冷冻保存和冷藏 可以优化切片，使新鲜组织的代谢活动 被保留。 通过对相关参数的系统评估 在冷冻和冷藏中，例如溶液的最佳组成， 冷冻速率、冷冻保护剂玻璃化条件、复温速率 及其对维持人体外源性代谢的影响 组织急需有关这种新的体外工具的一般知识 生化药理学将会出现。 外源性物质的稳定性 人类肝脏和肾脏切片培养过程中的代谢途径 准生理条件将告诉我们可以最大程度地预期什么 从这个体外系统。 由于该提案的重点是 整合的外源代谢，即 I 相和 I 相的紧密耦合 II 在完整的中间代谢控制条件下 蜂窝系统我们目前不会尝试确定数量， 个体的最大活性、合成率和降解率 酶，但将重点关注一些标准的复杂代谢模式 异生物质及其随着单个酶的脱落而发生的外观变化 或在文化过程中出现。

项目成果

Protein arylation precedes acetaminophen toxicity in a dynamic organ slice culture of mouse kidney.

在小鼠肾脏的动态器官切片培养物中，蛋白质芳基化先于对乙酰氨基酚毒性。

• DOI: 10.1006/faat.1996.0180
• 发表时间: 1996
• 期刊: Fundamental and applied toxicology : official journal of the Society of Toxicology
• 作者: Hoivik,DJ;Fisher,RL;Brendel,K;Gandolfi,AJ;Khairallah,EA;Cohen,SD
• 通讯作者: Cohen,SD